Comprehensive T-cell Characterization Assays

IFN-γ ELISPOT and cytometric bead array (CBA) assays were carried out as previously described [37 (link)] and intracellular cytokine staining and staining of cell surface molecules was carried out as previously described by Burgers et al., 2006 [47 (link)]. The following antibodies were used; anti-CD3⁺ Alexa 700, anti-CD4⁺ PE-Cy7, anti-CD8⁺ APC-Cy7, anti-CD62L APC, and anti-CD44 FITC). The cytokine antibodies were all PE-conjugated (0.2μg anti-TNF-PE, 0.06 μg anti-IL-2-PE, 0.06 μg anti-IFN-γ-PE) and were obtained from BD biosciences. The anti-CD4⁺ PE-Cy7, anti-CD8⁺ APC-Cy7 were obtained from BD biosciences. All the other antibodies were obtained from eBioscience.